Department of Internal Medicine 3 - Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU) and University Hospital Erlangen, Erlangen, Germany.
Translational Center Würzburg, Fraunhofer Translational Center Regenerative Therapies, Fraunhofer Institute for Silicate Research (ISC), Würzburg, Germany.
Ann Rheum Dis. 2019 Dec;78(12):1686-1692. doi: 10.1136/annrheumdis-2019-216108. Epub 2019 Sep 20.
Fibrosis is a complex pathophysiological process involving interplay between multiple cell types. Experimental modelling of fibrosis is essential for the understanding of its pathogenesis and for testing of putative antifibrotic drugs. However, most current models employ either phylogenetically distant species or rely on human cells cultured in an artificial environment. Here we evaluated the potential of vascularised in vitro human skin equivalents as a novel model of skin fibrosis and a platform for the evaluation of antifibrotic drugs.
Skin equivalents were assembled on a three-dimensional extracellular matrix by sequential seeding of endothelial cells, fibroblasts and keratinocytes. Fibrotic transformation on exposure to transforming growth factor-β (TGFβ) and response to treatment with nintedanib as an established antifibrotic agent were evaluated by quantitative polymerase chain reaction (qPCR), capillary Western immunoassay, immunostaining and histology.
Skin equivalents perfused at a physiological pressure formed a mature, polarised epidermis, a stratified dermis and a functional vessel system. Exposure of these models to TGFβ recapitulated key features of SSc skin with activation of TGFβ pathways, fibroblast to myofibroblast transition, increased release of collagen and excessive deposition of extracellular matrix. Treatment with the antifibrotic agent nintedanib ameliorated this fibrotic transformation.
Our data provide evidence that vascularised skin equivalents can replicate key features of fibrotic skin and may serve as a platform for evaluation of antifibrotic drugs in a pathophysiologically relevant human setting.
纤维化是一个涉及多种细胞类型相互作用的复杂病理生理过程。纤维化的实验模型对于理解其发病机制和测试潜在的抗纤维化药物至关重要。然而，大多数当前的模型采用的是系统发育上遥远的物种，或者依赖于在人工环境中培养的人类细胞。在这里，我们评估了血管化体外人皮肤等效物作为皮肤纤维化的新型模型以及评估抗纤维化药物的平台的潜力。
通过顺序接种内皮细胞、成纤维细胞和角质形成细胞，将皮肤等效物组装在三维细胞外基质上。通过定量聚合酶链反应（qPCR）、毛细管 Western 免疫测定、免疫染色和组织学评估在暴露于转化生长因子-β（TGFβ）和用尼达尼布（一种已建立的抗纤维化药物）治疗时的纤维化转化和反应。
在生理压力下灌注的皮肤等效物形成了成熟的、极化的表皮、分层的真皮和功能血管系统。这些模型暴露于 TGFβ 可再现 SSc 皮肤的关键特征，包括 TGFβ 途径的激活、成纤维细胞向肌成纤维细胞的转化、胶原释放增加和细胞外基质的过度沉积。用抗纤维化药物尼达尼布治疗可改善这种纤维化转化。
我们的数据提供了证据，表明血管化皮肤等效物可以复制纤维化皮肤的关键特征，并可作为在生理相关的人类环境中评估抗纤维化药物的平台。
